Some arterial dysfunction occurs due to narrowing of the arteries by fatty deposits or other vascular abnormalities. This may interfere with blood flow and/or prevent tissues and organs from being supplied with sufficient nutrients and oxygen.
Vascular disorders are common conditions and can severely compromise a patient's quality of life. Despite considerable advances in medical therapy and improvements in revascularization procedures for artery dysfunction, such as coronary artery graft, balloon angioplasty and stenting of the coronary vessels, a substantial proportion of patients suffer from artery dysfunction-derived disease.
Endothelial progenitor cells (EPCs) have been used to treat patients suffering from vascular diseases. In such severe cases, when drugs or direct revascularization procedures are not effective anymore, or cannot be used, alternative therapies are required. EPCs have been applied to ischemic tissue. EPCs have the ability to differentiate in order to form endothelium, the layer of cells forming blood vessels. These cells are involved in re-endothelialization, neovascularization, vasculogenesis and angiogenesis processes. The mechanisms by which implanted EPCs can become part of the healing process include self-repopulation, fusion with cells of the injured tissue and secretion of cytokines and growth factors. Repopulation of EPCs and their differentiation into mature endothelial cells enables their functions in re-endothelialization, neovascularization, vasculogenesis and angiogenesis processes. Recent evidence suggests that fusion of EPCs with cells of injured tissue enhances tissue function regeneration. Moreover, following secretion of cytokines and growth factors, EPCs can influence cellular survival of cells inherent to the tissue, and may help the mobilization of stem cells to the injured tissue.
A common ancestor cell, the hemangioblast, gives rise to both endothelial and hematopoietic (blood cell) precursors. This ancestor cell differentiates into hematopoietic stem cells and angioblasts, which are mesodermal precursor cells, differentiating into endothelial precursors. These cells have the capacity to proliferate, migrate, and differentiate into endothelial cells, but have not yet acquired specific mature endothelial markers. Following commitment to the endothelial lineage, angioblasts assemble into a primitive vascular plexus of veins and arteries, in a process called vasculogenesis. This primitive vasculature is subsequently refined into a functional network by angiogenesis and by remodeling and arteriogenesis of newly formed vessels. EPCs have been shown to mobilize (i.e., migrate in increased numbers from the bone marrow (BM) into the circulation) in patients with vascular trauma or Acute Myocardial Infarction (AMI) (See, for example, the following two articles which are incorporated herein by reference: (a) Gill, M., S. Dias, et al. (2001), “Vascular trauma induces rapid but transient mobilization of VEGFR2(+)AC133(+) endothelial precursor cells,” Circ Res 88(2): 167-74; and (b) Shintani, S., T. Murohara, et al. (2001), “Mobilization of endothelial progenitor cells in patients with acute myocardial infarction,” Circulation 103(23): 2776-9.) In general, the use of EPCs aims to promote the formation of natural bypasses within the ischemic or scarred tissue and thus alleviate the clinical condition of these patients.
Numerous animal experiments and clinical trials have investigated the potential of this therapy to augment blood flow and yield an associated alleviation of ischemic symptoms, as manifested by a patient's improvement in physical functioning.
Various sources for autologous EPCs for transplantation have been described, including stem cells aspirated directly from the bone marrow (BM), and BM-derived peripheral blood stem cells.
Progenitor cells, or stem cells, include bone marrow cells that can multiply, migrate and differentiate into a wide variety of cell types. Bone marrow hematopoietic stem cells are characterized as being “CD34 positive” (CD34+), i.e., expressing the CD34 marker.
It is assumed that the plasticity of well-defined populations of hematopoietic progenitors allows them to trans-differentiate in response to the environmental cues present in the target organ, and, more specifically, to convert into endothelial cells.
Transplantation of bone marrow is clinically appealing because of the relative simplicity of the medical procedure. It entails aspiration of bone marrow from the iliac crest and immediate re-injection of the aspirate or selected cells into the post-infarction scar. Nevertheless, the procedure is invasive and must be done under anesthesia.
The first evidence indicating the presence of EPCs in the adult circulation was obtained when mononuclear blood cells from healthy human volunteers were shown to acquire an endothelial cell-like phenotype in vitro and to incorporate into capillaries in vivo. (See Asahara, T., T. Murohara, et al. (1997), “Isolation of putative progenitor endothelial cells for angiogenesis,” Science 275(5302): 964-7, which is incorporated herein by reference.) These putative EPCs were characterized via expression of CD34 and vascular endothelial growth factor receptor-2 (VEGFR-2/KDR), two antigens shared by embryonic endothelial progenitors, and hematopoietic stem cells (HSCs). In addition to CD34, early hematopoietic progenitor cells express CD133 (AC133), which is not expressed after differentiation. Currently, the widely accepted definition of EPCs in circulation is, for practical purposes, CD34+/VEGFR-2+ or CD133+/VEGFR-2+ cells.
Peripheral blood EPCs can be obtained from blood of untreated patients or from patients treated in order to augment EPC mobilization using cytokines such as granulocyte-colony stimulating factor (G-CSF), granulocyte monocyte colony-stimulating factor (GM-CSF), vascular endothelial growth factor (VEGF), and fibroblast growth factor (FGF). The mobilization treatments are typically avoided in patients who suffer from hematological and arterial derived disorders. Treatments such as HMG CoA reductase inhibitors (statins) have also been reported to elevate numbers of EPCs in circulation. See, for example:                1. Dimmeler S., et al. (2001), “HMG-CoA reductase inhibitors (statins) increase endothelial progenitor cells via the PI 3-kinase/Akt pathway,” J. Clin. Invest. 108: 391-397.        2. Hyun-Jae, Hyo-Soo Kim, et al. (2003), “Effects of intracoronary infusion of peripheral blood stem-cells mobilized with granulocyte-colony stimulating factor on left ventricular systolic function and restenosis after coronary stenting in myocardial infraction: the MAGIC cell randomized clinical trial,” The Lancet 363: 751-756.        3. Brigit Assmus, Volker Schachinger et al., (2002), “Transplantation of progenitor cells and regeneration enhancement in acute myocardial infraction (TOPCARE-AMI),” Circulation 106: 3009-3017.        4. Alexandra Aicher, Winfreid Brenner, et al., (2003), “Assessment of the tissue distribution of transplanted human endothelial progenitor cells by radioactive labeling,” Circulation 107: 2134-2139.        
Each of these articles is incorporated herein by reference.
The procedure for removing peripheral blood is simpler and more convenient for the patient than BM removal. The fact that EPCs can be isolated from peripheral blood is an additional important factor in the choice of using these cells for therapy. The isolation of progenitor cells from BM, which contains many more cell types, is technically more challenging, as well.
Results from EPC and BMC treatments show improved cardiac function, greater capillary density, marked increase in number of collateral vessels, improvement of echocardiographic left ventricular ejection fraction, decrease in ischemic area scarring and prevention of cardiomyocyte apoptosis in rat models of myocardial infarction. Furthermore, improved blood flow and capillary density and reduced rate of limb loss in hindlimb was shown in an ischemia model in nude mice.
The following articles, which are incorporated herein by reference, describe techniques which may be used in combination with techniques described herein:                (1) Kalka, C., H. Masuda, et al. (2000). “Vascular endothelial growth factor (165) gene transfer augments circulating endothelial progenitor cells in human subjects.” Circ Res 86(12): 1198-202.        (2) Kawamoto, A., H. C. Gwon, et al. (2001). “Therapeutic potential of ex vivo expanded endothelial progenitor cells for myocardial ischemia.” Circulation 103(5): 634-7.        (3) Kawamoto, A., T. Tkebuchava, et al. (2003). “Intramyocardial transplantation of autologous endothelial progenitor cells for therapeutic neovascularization of myocardial ischemia.” Circulation 107(3): 461-8.        (4) Kamihata, H., H. Matsubara, et al. (2001). “Implantation of bone marrow mononuclear cells into ischemic myocardium enhances collateral perfusion and regional function via side supply of angioblasts, angiogenic ligands, and cytokines.” Circulation 104(9): 1046-52.        (5) Kocher, A. A., M. D. Schuster, et al. (2001). “Neovascularization of ischemic myocardium by human bone-marrow-derived angioblasts prevents cardiomyocyte apoptosis, reduces remodeling and improves cardiac function.” Nat Med 7(4): 430-6.        
The following articles and book chapter, which are also incorporated herein by reference, describe techniques which may be used in combination with techniques described herein:                Flammera J et al. (2002). “The impact of ocular blood flow in glaucoma.” Progress in Retinal and Eye Research 21:359-393.        Zarbin M A (2004). “Current concepts in the pathogenesis of age-related macular degeneration.” Arch Opthalmol. 122(4):598-614.        Frank R N (2004). “Diabetic retinopathy.” N Engl J Med 350:48-58.        Singleton J R (2003). “Microvascular complications of impaired glucose tolerance.” Diabetes 52:2867-2873.        Bahlmann F H et. (2004). “Erythropoietin regulates endothelial progenitor cells.” Blood 103(3):921-6.        Greenfield, Ed. (2001). “Surgery: scientific principles and practice.” Lippincot: Philadelphia, chapter 107.        Kouwenhoven E A et al. (2000). “Etiology and pathophysiology of chronic transplant dysfunction.” Transplant Internat. 13(6):385-401.        Browne E Z et al. (1986). “Complications of skin grafts and pedicle flaps.” Hand Clin. 2:353-9.        Chen et al. (1991). “Four types of venous flaps for wound coverage: a clinical appraisal.” J. Trauma 31(9):1286-93.        Beatrice et al. (2004) Dermatol. Surg. 30(3):399.        Ferretti et al. (2003). “Angiogenesis and nerve regeneration in a model of human skin equivalent transplant.” Life Sci. 73:1985-94.        Schechner et al. (2003). “Engraftment of a vascularized human skin equivalent.” FASEB J. 17(15):2250-60.        
Research has been carried out in humans during the last few years to examine the potential benefits of using EPCs and other bone marrow derived cells to treat myocardial disorders. Recent studies demonstrate that implantation of autologous progenitor cells after Acute Myocardial Infarction appears to limit post-infarction damage. The following clinical trials focus on the studies that assessed the safety and efficacy of bone marrow-derived or blood-derived cells administered in patients with cardiac disorders.
Perin et al. carried out a clinical trial which included 21 patients (14 in the treatment group, 7 in the control group) who received transendocardial injections of autologous mononuclear BMCs. (See Perin, E. C., H. F. Dohmann, et al. (2003), “Transendocardial, autologous bone marrow cell transplantation for severe, chronic ischemic heart failure,” Circulation 107(18): 2294-302, which is incorporated herein by reference.) At 4 months, there was an improvement in ejection fraction, a reduction in end-systolic volume, and significant mechanical improvement of the injected segments in the treated patients.
Another group injected autologous EPCs into the infarct border zone in six patients who had suffered from myocardial infarction and undergone coronary artery bypass grafting. Three to nine months after surgery, all patients were alive and well, and global left-ventricular function was enhanced in four patients. All six patients reported a notable improvement in exercise capacity. Myocardial perfusion scans were reported to have improved strikingly by qualitative analysis in five of six patients. The results of this study indicate that implantation of EPCs to the heart probably induces angiogenesis, thus improving perfusion of the infarcted myocardium. (See Stamm, C., B. Westphal, et al. (2003), “Autologous bone-marrow stem-cell transplantation for myocardial regeneration,” Lancet 361(9351): 45-6, which is incorporated herein by reference.)
The Transplantation of Progenitor Cells and Regeneration Enhancement in Acute Myocardial Infarction (TOPCARE-AMI) study involved the delivery of circulating endothelial progenitor cells or bone marrow cells directly into coronary arteries after the infarction in patients with reperfused acute myocardial infarction (See Assmus, B., V. 15, Schachinger, et al. (2002), “Transplantation of Progenitor Cells and Regeneration Enhancement in Acute Myocardial Infarction (TOPCARE-AMI),” Circulation 106(24): 3009-17, which is incorporated herein by reference.) In the first 20 patients, 11 received EPCs and 9 received BMCs. At 4 months, transplantation of progenitor cells resulted in a significant increase in global left ventricular ejection fraction, echocardiography revealed improved regional wall motion in the infarct zone, reduced end-systolic left ventricular volumes, and increased myocardial viability in the infarct zone compared with a nonrandomized, matched reference group. There were no adverse events of treatment in any of the patients such arrhythmias, or increase in creatine kinase and troponin. There was no difference between BM- and peripheral blood-derived cells.
A study performed by a team led by Amit Patel of University of Pittsburgh, involved 20 patients with severe heart failure out of which 10 were injected into the coronary vessels with BM derived EPCs. At one-, three- and six-month follow-up, the ejection fraction rates for the stem cell patients were significantly improved compared to the other patients. (See Abstract from American Association for Thoracic Surgery, Toronto, May 2004).
The following articles, which are incorporated herein by reference, may also be of interest:                J. Folkman, Y. Shing, J. Biol. Chem. 267, 10931 (1992)        W. Brugger, S. Heimfeld, R. J. Berenson, R. Mertelsmann, L. Kanz, N. Engl J. Med. 333, 283 (1995)        F. Katz, R. W. Tindle, D. R. Sutherland, M. D. Greaves, Leuk. Res. 9, 191 (1985)        R. G. Andrews, J. W. Singer, I. D. Bernstein, Blood 67, 842 (1986)        B. I. Terman, M. Dougher-Vermazen, M. E. Carrion, D. Dimitrov, D. C. Armellino, et al, Biochem. Biophys. Res. Commun. 187, 1579 (1992)        B. Millauer, S. Wizigmann-Voos, H. Schnurch, R. Martinez, N. P. H. Moller, et al, Cell 72, 835 (1993)        J. C. Voyta, D. P. Via, C. E. Butterfield, B. R. Zetter, J. Cell Biol. 99, 2034 (1984)        P. J. Newman, M. C. Berndt, J. Gorski, G. C. White, S. Lyman, et al, Science 247, 1219 (1990)        T. N. Sato, Y. Tozawa, U. Deutsch, K. Wolburg-Buchholz, Y. Fujiwara, et al, Nature 376, 70 (1995)        H. Schnurch, W. Risau, Development 119, 957 (1993)        J. L. Liesveld, K. E. Frediani, A. W. Harbol, J. F. DiPersio, C. N. Abboud, Leukemia 8, 2111 (1994).        S. Takeshita, L. P. Zheng, E. Brogi, M. Kearney, L. Q. Pu, et al, J. Clin. Invest. 93, 662 (1994)        R. Baffour, J. Berman, J. L. Garb, S. W. Rhee, J. Kaufman, et al, J. Vase. Surg. 16, 181 (1992)        J. M. Isner, A. Pieczek, R. Schainfeld, R. Blair, L. Haley, et al, Lancet 348, 370 (1996)        Y. Sato, K. Okamura, A. Morimoto, R. Hamanaka, K. Hamanaguchi, et al, Exp. Cell Res. 204, 223 (1993)        
The following articles, which are also incorporated herein by reference, may also be of interest:                Badorff, C., R. P. Brandes, et al. (2003). “Transdifferentiation of blood-derived human adult endothelial progenitor cells into functionally active cardiomyocytes.” Circulation 107(7): 1024-32.        Bhattacharya, V., P. A. McSweeney, et al. (2000). “Enhanced endothelialization and microvessel formation in polyester grafts seeded with CD34(+) bone marrow cells.” Blood 95(2): 581-5.        Grant, M. B., W. S. May, et al. (2002). “Adult hematopoietic stem cells provide functional hemangioblast activity during retinal neovascularization.” Nat Med 8(6): 607-12.        Hirata, K., T. S. Li, et al. (2003). “Autologous bone marrow cell implantation as therapeutic angiogenesis for ischemic hindlimb in diabetic rat model.” Am J Physiol Heart Circ Physiol 284(1): H66-70.        Ikenaga, S., K. Hamano, et al. (2001). “Autologous bone marrow implantation induced angiogenesis and improved deteriorated exercise capacity in a rat ischemic hindlimb model.” J Surg Res 96(2): 277-83.        Kalka, C., H. Masuda, et al. (2000). “Transplantation of ex vivo expanded endothelial progenitor cells for therapeutic neovascularization.” Proc Natl Acad Sci USA 97(7): 3422-7.        Kaushal, S., G. E. Amiel, et al. (2001). “Functional small-diameter neovessels created using endothelial progenitor cells expanded ex vivo.” Nat Med 7(9): 1035-40.        Kornowski, R., M. B. Leon, et al. (2000). “Electromagnetic guidance for catheter-based transendocardial injection: a platform for intramyocardial angiogenesis therapy. Results in normal and ischemic porcine models.” J Am Coll Cardiol 35(4): 1031-9.        Li, R. K., Z. Q. Jia, et al. (1996). “Cardiomyocyte transplantation improves heart function.” Ann Thorac Surg 62(3): 654-60; discussion 660-1.        Rajnoch, C., J. C. Chachques, et al. (2001). “Cellular therapy reverses myocardial dysfunction.” J Thorac Cardiovasc Surg 121(5): 871-8.        Schatteman, G. C., H. D. Hanlon, et al. (2000). “Blood-derived angioblasts accelerate blood-flow restoration in diabetic mice.” J Clin Invest 106(4): 571-8.        Shintani, S., T. Murohara, et al. (2001). “Augmentation of postnatal neovascularization with autologous bone marrow transplantation.” Circulation 103(6): 897-903.        Strauer, B. E., M. Brehm, et al. (2002). “Repair of infarcted myocardium by autologous intracoronary mononuclear bone marrow cell transplantation in humans.” Circulation 106(15): 1913-8.        Taylor, D. A., B. Z. Atkins, et al. (1998). “Regenerating functional myocardium: improved performance after skeletal myoblast transplantation.” Nat Med 4(8): 929-33.        Thompson, C. A., B. A. Nasseri, et al. (2003). “Percutaneous transvenous cellular cardiomyoplasty. A novel nonsurgical approach for myocardial cell transplantation.” J Am Coll Cardiol 41(11): 1964-71.        Tomita, S., R. K. Li, et al. (1999). “Autologous transplantation of bone marrow cells improves damaged heart function.” Circulation 100(19 Suppl): 11247-56.        Tomita, S., D. A. Mickle, et al. (2002). “Improved heart function with myogenesis and angiogenesis after autologous porcine bone marrow stromal cell transplantation.” J Thorac Cardiovasc Surg 123(6): 1132-40.        Wang et al. (2004). “Rosiglitazone facilitates angiogenic progenitor cell differentiation toward endothelial lineage: a new paradigm in glitazone pleiotropy.” Circulation 109(11): 1392-400.        Rupp et al. (2004). “Statin therapy in patients with coronary artery disease improves the impaired endothelial progenitor cell differentiation into cardiomyogenic cells.” Basic Res Cardiol. 99(1): 61-8.        Quirici et al. (2001). “Differentiation and expansion of endothelial cells from human bone marrow CD133(+) cells.” Br J. Haematol. 115(1): 186-94.        Di Stefano et al. (2002) “Different growth conditions for peripheral blood endothelial progenitors.” Cardiovasc Radiat Med. 3(3-4): 172-5.        Akita et al. (2003). “Hypoxic preconditioning augments efficacy of human endothelial progenitor cells for therapeutic neovascularization.” Lab Invest. 83(1): 65-73.        Wang et al. (2004). “Mechanical, cellular, and molecular factors interact to modulate circulating endothelial cell progenitors.” Am J Physiol Heart Circ Physiol. 286(5): H1985-93.        Bahlmann et al. (2003). “Endothelial progenitor cell proliferation and differentiation is regulated by erythropoietin.” Kidney Int. 64(5): 1648-52.        Heeschen et al. (2003). “Erythropoietin is a potent physiologic stimulus for endothelial progenitor cell mobilization.” Blood. 102(4): 1340-6.        Verma et al. (2004). “C-reactive protein attenuates endothelial progenitor cell survival, differentiation, and function: Further evidence of a mechanistic link between C-reactive protein and cardiovascular disease.” Circulation. 109(17): 2058-67.        
U.S. Pat. Nos. 5,980,887, 6,569,428, and 6,676,937 to Isner et al., which are incorporated herein by reference, generally describe pharmaceutical products including EC progenitors for use in methods for regulating angiogenesis, i.e., for enhancing or inhibiting blood vessel formation, in a selected patient and in some preferred embodiments for targeting an angiogenesis modulator to specific locations. For example, the EC progenitors can be used to enhance angiogenesis or to deliver an angiogenesis modulator, e.g., anti- or pro-angiogenic agents, respectively to sites of pathologic or utilitarian angiogenesis. Additionally, in another embodiment, EC progenitors can be used to induce re-endothelialization of an injured blood vessel, and thus reduce restenosis by indirectly inhibiting smooth muscle cell proliferation.
U.S. Pat. No. 5,541,103 to Kanz et al., which is incorporated herein by reference, describes high-dose chemotherapy treatments for patients suffering from certain types of cancer. In order to facilitate recovery, a process for the ex vivo expansion of peripheral blood progenitor cells is described, wherein CD34+ cells are enriched and cultivated in a medium comprising IL-1, IL-3, IL-6, EPO and SCF. The ex vivo expanded peripheral blood progenitor cells can be administered to cancer patients after chemotherapy.
US Patent Application Publication 2003/0199464 to Itescu, which is incorporated herein by reference, describes a method for treating a disorder of a subject's heart involving loss of cardiomyocytes. The method includes administering to the subject an amount of an agent described as being effective to cause cardiomyocyte proliferation within the subject's heart so as to thereby treat the disorder. In an embodiment, the agent is human endothelial progenitor cells. The application also describes methods for determining the susceptibility of a cardiomyocyte in a subject to apoptosis.
PCT Patent Publication WO 01/94420 to Itescu, which is incorporated herein by reference, describes a method of stimulating vasculogenesis of myocardial infarct damaged tissue in a subject comprising: (a) removing stem cells from a location in the subject; (b) recovering endothelial progenitor cells from the stem cells; (c) introducing the endothelial progenitor cells from step (b) into a different location in the subject such that the precursors migrate to and stimulate revascularization of the tissue. The stem cells may be removed directly or by mobilization. The endothelial progenitor cells may be expanded before introduction into the subject. A method of inducing angiogenesis in peri-infarct tissue is described. A method is also described for selectively increasing the trafficking of human bone marrow-derived endothelial cell precursors to the site of tissue damaged by ischemic injury, which comprises: (a) administering endothelial progenitor cells to a subject; (b) administering chemokines to the subject so as to thereby attract endothelial cell precursors to the ischemic tissue. A method is also described for stimulating vasculogenesis or angiogenesis of myocardial infarct damaged tissue in a subject comprising injecting allogeneic stem cells into a subject. A method is also described for improving myocardial function in a subject that has suffered a myocardial infarct comprising any of the instant methods. A method is also described for improving myocardial function in a subject who has suffered a myocardial infarct comprising injecting G-CSF or anti-CXCR4 antibody into the subject in order to mobilize endothelial progenitor cells.
U.S. Pat. No. 5,199,942 to Gillis, describes a method for autologous hematopoietic cell transplantation of patients receiving cytoreductive therapy, including: (1) obtaining hematopoietic progenitor cells from bone marrow or peripheral blood from a patient prior to cytoreductive therapy; (2) expanding the hematopoietic progenitor cells ex vivo with an ex vivo growth factor selected from the group consisting of interleukin-3 (IL-3), steel factor (SF), granulocyte macrophage-colony stimulating factor (GM-CSF), interleukin-1 (IL-1), GM-CSF/IL-3 fusion proteins and combinations thereof, to provide a cellular preparation comprising an expanded population of progenitor cells; and (3) administering the cellular preparation to the patient concurrently with or following cytoreductive therapy. The method optionally includes a preliminary treatment with a recruitment growth factor to recruit hematopoietic progenitor cells into peripheral blood and a subsequent treatment with an engraftment growth factor to facilitate engraftment and proliferation of hematopoietic progenitor cells administered in the cellular preparation. The patent also describes a hematopoietic progenitor cell expansion media composition comprising cell media, an ex vivo growth factor, and autologous serum.
U.S. Pat. No. 4,656,130 to Shoshan, which is incorporated herein by reference, describes a collagen coated cell growth plate that includes a substrate coated with a storage stable coating of collagen fibrils. The method of preparing the collagen coated cell growth plates comprises dispensing biologically active collagen fibrils suspended in distilled water onto a tissue culture dish. Thereafter, the dish containing the collagen fibril suspension is placed in a laminar flow hood provided with a sterile air stream and ultraviolet light. The fibrils sediment and adhere to the bottom of the dish, the water evaporates in the sterile air stream and is removed in the laminar flow hood exhaust, and the ultraviolet light ensures that the resulting thin layer of collagen fibrils is sterile and ready for the inoculation of living cells. The method is described as yielding a convenient precoated cell growth plate which can maintain reasonable shelf life when kept at room temperature without any significant decrease in cell growth support properties.
U.S. Pat. No. 5,932,473 to Swiderek et al., which is incorporated herein by reference, describes a cell culture substrate that is coated with a composition containing a cell adhesion promoter in a salt solution. A substrate such as plastic, glass or microporous fibers is coated with a composition containing about 5-1000 ug/ml of poly-D-lysine in an 0.005-0.5 M citrate or sulfate salt solution, in order to provide about 50-500 ul of the composition per cm2 of substrate. The coated substrate is rinsed to remove extraneous materials, and dried to obtain a coated substrate having increased shelf-life and/or stability. The coated substrate may be sterilized by rinsing with a sterilizing medium such as ethanol.
U.S. Pat. No. 6,040,182 to Septak, which is incorporated herein by reference, describes methods and materials for the facilitation of high-protein-binding capability on tissue culture-treated plastic surfaces, such as, for example, polystyrene assay plates.
U.S. Pat. No. 4,450,231 to Ozkan, which is incorporated herein by reference, describes an immunoassay of a specimen of a serum or the like to determine immune complexes. A method is described which includes producing on a plastic base a layer of a non-proteinaceous, non-ionic polymer which will adhere to the plastic base and has the capability of absorbing immune complexes of the specimen, placing a specimen on the layer and treating the layer to produce an indication of the amount of immune complexes. The polymer may be polyethylene glycol, dextran, polyvinyl chloride, a polymeric polyol or an adduct of polyethylene glycol. A product for use in such an assay is a plate having wells or a test tube formed of plastic, polystyrene and polyvinyl chloride being preferred, with a layer of such non-proteinaceous, non-ionic layer on the plate wells or the cavity of the test tube.
US Patent Application Publication 2003/0229393 to Kutryk et al., which is incorporated herein by reference, describes compositions and methods for producing a medical device such as a stent, a stent graft, a synthetic vascular graft, or heart valves, which are coated with a biocompatible matrix which incorporates antibodies, antibody fragments, or small molecules, which recognize, bind to and/or interact with a progenitor cell surface antigen to immobilize the cells at the surface of the device. The coating on the device can also contain a compound or growth factor for promoting the progenitor endothelial cell to accelerate adherence, growth and differentiation of the bound cells into mature and functional endothelial cells on the surface of the device to prevent intimal hyperplasia. Methods for preparing such medical devices, compositions, and methods for treating a mammal with vascular disease such as restenosis, atherosclerosis or other types of vessel obstructions are described.